Contrast-induced nephropathy in an animal model: Evaluation of novel biomarkers in blood and tissue samples

Charalampos Mamoulakis; Irene Fragkiadoulaki; Phaedra Karkala; Georgios Georgiadis; Ioannis-Erineos Zisis; Polychronis Stivaktakis; Alexandra Kalogeraki; Ioannis Tsiaoussis; Tatyana Burykina; George Lazopoulos; Konstantinos Tsarouhas; Dimitrios Kouretas; Aristides Tsatsakis

doi:10.1016/j.toxrep.2019.04.007

Contrast-induced nephropathy in an animal model: Evaluation of novel biomarkers in blood and tissue samples

Toxicol Rep. 2019 May 2:6:395-400. doi: 10.1016/j.toxrep.2019.04.007. eCollection 2019.

Authors

Charalampos Mamoulakis¹, Irene Fragkiadoulaki^{1

2}, Phaedra Karkala², Georgios Georgiadis¹, Ioannis-Erineos Zisis^{1

2}, Polychronis Stivaktakis², Alexandra Kalogeraki³, Ioannis Tsiaoussis⁴, Tatyana Burykina⁵, George Lazopoulos⁶, Konstantinos Tsarouhas⁷, Dimitrios Kouretas⁸, Aristides Tsatsakis^{2

5}

Affiliations

¹ Department of Urology, University General Hospital of Heraklion, University of Crete, Medical School, Heraklion, Crete, Greece.
² Department of Forensic Sciences and Toxicology, Faculty of Medicine, University of Crete, Heraklion, 71003, Greece.
³ Department of Pathology-Cytopathology, University General Hospital of Heraklion, University of Crete, Medical School, Heraklion, Crete, Greece.
⁴ Laboratory of Anatomy-Histology-Embryology, University of Crete, Medical School, Heraklion, Crete, Greece.
⁵ Department of Analytical and Forensic Medical Toxicology, Sechenov University, 2-4 Bolshaya Pirogovskaya st., 119991, Moscow, Russia.
⁶ Department of Cardiothoracic Surgery, University General Hospital of Heraklion, University of Crete, Medical School, Heraklion, Crete, Greece.
⁷ Department of Cardiology, University General Hospital of Larisa, Larissa, Greece.
⁸ Department of Biochemistry and Biotechnology, University of Thessaly, 41500, Larissa, Greece.

Abstract

Identification of novel biomarkers of contrast-induced nephropathy (CIN) that may more accurately detect renal function changes; reflect kidney damage; assist monitoring; and elucidate pathophysiology attract considerable scientific attention nowadays. To evaluate novel biomarkers of nephrotoxicity in blood/tissue samples of a CIN model, 10 New Zealand white rabbits were divided into group 1 (n = 5; iopromide) and group 2 (n = 5; control). Blood was drawn at 0 h (immediately), 24 h and 48 h after contrast medium (CM) administration. Animals were euthanized at 48 h and kidneys were removed. Serum creatinine (sCr)/symmetric-asymmetric dimethylarginine (SDMA-ADMA) levels were measured. CM genotoxic/cytotoxic effect was investigated 48 h post-CM exposure using micronucleus assay in lymphocytes. Cytological examination was conducted using touch preparation technique (TPT). All animals in group 1 developed CIN: mean sCr levels increased by 68.2% within 48 h. Significant SDMA-ADMA level elevation was observed at 0 h and 24 h with insignificant drop at 48 h in group 1, remaining normal in group 2 at all time-points. Significant increase in bi-nucleated cells with micronuclei and micronuclei frequency was detected in group 1. Cytokinesis block proliferation index was reduced insignificantly in group 1. TPT revealed degenerative lesions/inflammation, cell degeneration, abnormal uterine tubular casts and rubella in kidneys of all animals in group 1. Group 2 presented normal cells.

Keywords: ADMA, asymmetric dimethylarginine; AKI, acute kidney injury; ANOVA, analysis of variance; ARRIVE, animal research: reporting of in vivo experiments; AVMA, American Veterinary Medical Association; Animal; Asymmetric dimethylarginine; BNMN, Bi-nucleated cells with micronuclei; CBPI, cytokinesis block proliferation index; CIN, contrast-induced nephropathy; CKD, chronic kidney disease; CM, contrast medium; Contrast media; ESI, electrospray ionization; GFR, glomerular filtration rate; Iopromide; KIM-1, kidney injury molecule-1; Kidney; LC–MS, liquid chromatography mass spectrometry; MN, micronuclei; Models; NGAL, meutrophil gelatinase–associated lipocalin; NO, nitric oxide; Nephropathy; Nephrotoxicity; OECD, Organisation for Economic Co-operation and Development; RBF, renal blood flow; ROS, reactive oxygen species; SCR, serum creatinine; SD, standard deviation; SDMA, symmetric dimethylarginine; Symmetric dimethylarginine; TPT, touch preparation technique.